Treatment of discs large homolog (DLG) related diseases by inhibition of natural antisense transcript to DLG

• US 8,980,860 B2
• Filed: 07/13/2011
• Issued: 03/17/2015
• Est. Priority Date: 07/14/2010
• Status: Active Grant

• Current Assignee: CURNA, Inc. (Opko Health, Inc.)
• Original Assignee: CURNA, Inc. (Opko Health, Inc.)
• Inventors: Collard, Joseph, Khorkova Sherman, Olga, Coito, Carlos
• Primary Examiner(s): GIBBS, TERRA C
• Application Number:
  US13/810,019
• Publication Number:
  US 20130143946A1
• Time in Patent Office: 1,343 Days
• Field of Search: 514/44, 536/24.5
• US Class Current: 514/44.A
• CPC Class Codes: A61K 31/7088 Compounds having three or more nucleosides or nucleotides
• CPC Class Codes: A61P 13/00 Drugs for disorders of the urinary system
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: A61P 25/18 Antipsychotics, i.e. neuroleptics; Drugs for mania or schizophrenia
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 31/18 for HIV
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 2310/111 spanning the whole gene, or a large part of it
• CPC Class Codes: C12N 2310/113 targeting other non-coding nucleic acids, e.g. antagomirs
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2330/10 naturally occurring
• CPC Class Codes: C12Q 1/6813 Hybridisation assays

Abstract

The present invention relates to antisense oligonucleotides that modulate the expression of and/or function of Discs large homolog (DLG), in particular, by targeting natural antisense polynucleotides of Discs large homolog (DLG). The invention also relates to the identification of these antisense oligonucleotides and their use in treating diseases and disorders associated with the expression of DLG.

Claims

1. A method of upregulating a function of and/or the expression of a Discs large homolog (DLG) polynucleotide in a mammal comprising:

contacting said mammal with at least one antisense oligonucleotide 12 to 30 nucleotides in length wherein said at least one oligonucleotide has at least 80% sequence identity to a reverse complement of a natural antisense of a Discs large homolog (DLG) polynucleotide;

thereby upregulating a function of and/or the expression of the Discs large homolog (DLG) polynucleotide.

2. A method of upregulating a function of and/or the expression of a Discs large homolog 1(DLG 1) polynucleotide in a mammal according to claim 1 comprising:

contacting said mammal with at least one antisense oligonucleotide 12 to 30 nucleotides in length wherein said at least one oligonucleotide has at least 80% sequence identity to a reverse complement of a polynucleotide comprising 12 to 30 nucleotides within the natural antisense transcript nucleotides 1 to 1706 of SEQ ID NO;

2;

thereby upregulating a function of and/or the expression of the Discs large homolog 1 (DLG

     1) polynucleotide.

14. The method of claim 1, wherein the at least one oligonucleotide comprises at least one oligonucleotide sequences set forth as SEQ ID NOS:

3 to 12.

3. A method of upregulating a function of and/or the expression of a Discs large homolog (DLG) polynucleotide in mammalian cells or tissues in vivo or in vitro comprising:

contacting said cells or tissues with at least one antisense oligonucleotide 12 to 30 nucleotides in length wherein said oligonucleotide has at least 90% sequence identity to a sense RNA transcribed from the Discs large homolog (DLG) polynucleotide;

thereby upregulating a function of and/or the expression of the Discs large homolog (DLG) polynucleotide in mammalian cells or tissues in vivo or in vitro.

4. A method of upregulating a function of and/or the expression of a Discs large homolog 1 (DLG 1) polynucleotide in mammalian cells or tissues according to claim 3 comprising:

contacting said mammalian cells or tissues with at least one antisense oligonucleotide 12 to 28 nucleotides in length wherein said at least one oligonucleotide has at least 80% sequence identity to a reverse complement of a polynucleotide comprising 12 to 28 consecutive nucleotides within the natural antisense transcript nucleotides 1 to 1706 of SEQ ID NO;

2;

thereby upregulating a function of and/or the expression of the Discs large homolog 1 (DLG

     1) polynucleotide.

5. A method of upregulating a function of and/or the expression of a Discs large homolog (DLG) polynucleotide having SEQ ID NO:

1 in a mammalian system comprising;

contacting said system with at least one antisense oligonucleotide of 12 to 28 nucleotides in length that targets and specifically hybridizes to a region of a natural antisense polynucleotide of the Discs large homolog (DLG) polynucleotide;

thereby upregulating a function of and/or the expression of the Discs large homolog (DLG) polynucleotide.

6. The method of claim 5, wherein a function of and/or the expression of the Discs large homolog (DLG) is increased in v'"'"'o or in vitro with respect to a control.

7. The method of claim 5, wherein the at least one antisense oligonucleotide targets a natural antisense sequence of a Discs large homolog 1 (DLG 1) polynucleotide having SEQ ID NO:

2.

8. The method of claim 5, wherein the at least one antisense oligonucleotide targets a natural antisense polynucleotide antisense to coding and/or non-coding nucleic acid sequences of a Discs large homolog 1<

(DLG

     1) polynucleotide.

9. The method of claim 5, wherein the at least one antisense oligonucleotide targets a natural antisense polynucleotide having overlapping and/or non-overlapping sequences of a Discs large homolog 1 (DLG 1) polynucleotide.

10. The method of claim 5, wherein the at least one antisense oligonucleotide comprises one or more modifications selected from:

at least one modified sugar moiety, at least one modified internucleoside linkage, at least one modified nucleotide, and combinations thereof.

11. The method of claim 10, wherein the one or more modifications comprise at least one modified sugar moiety selected from:

a 2′

-0-methoxyethyl modified sugar moiety, a 2′

-methoxy modified sugar moiety, a 2′

-0-alkyl modified sugar moiety, a bicyclic sugar moiety, and combinations thereof.

12. The method of claim 10, wherein the one or more modifications comprise at least one modified internucleoside linkage selected from:

a phosphorothioate, alkylphosphonate, phosphorodithioate, alkylphosphonothioate, phosphoramidate, carbamate, carbonate, phosphate triester, acetamidate, carboxymethyl ester, and combinations thereof.

13. The method of claim 10, wherein the one or more modifications comprise at least one modified nucleotide selected from:

a peptide nucleic acid (PNA), a locked nucleic acid (LNA), an arabino-nucleic acid (FANA), an analogue, a derivative, and combinations thereof.

15. A method of upregulating a function of and/or the expression of a Discs large homolog 1 (DLG 1) gene in mammalian cells or tissues in vivo or in vitro comprising:

contacting said cells or tissues with at least one short interfering RNA (siRNA) oligonucleotide 19 to 30 nucleotides in length, said at least one siRNA oligonucleotide being specific for a natural antisense polynucleotide of a Discs large homolog 1 (DLG

     1) polynucleotide, wherein said at least one siRNA oligonucleotide has at least 80% sequence complementarity to said natural antisense polynucleotide of the Discs large homolog 1 (DLG1) polynucleotide; and

, upregulating a function of and/or the expression of Discs large homolog 1 (DLG1) in mammalian cells or tissues in vivo or in vitro.

16. The method of claim 15, wherein said oligonucleotide has at least 90% sequence complementarity to said natural antisense polynucleotide of the Discs large homolog 1 (DLG1) polynucleotide.

17. A method of upregulating a function of and/or the expression of Discs large homolog 1 (DLG 1) in mammalian cells or tissues in vivo or in vitro comprising:

contacting said cells or tissues with at least one antisense oligonucleotide of about 12 to 30 nucleotides in length specific for noncoding and/or coding sequences of a natural antisense strand of a Discs large homolog 1 (DLG

     1) polynucleotide wherein said at least one antisense oligonucleotide has at least 80% sequence identity to at least one nucleotide sequence set forth as SEQ ID NO;

1; and

, upregulating the function and or expression of the Discs large homolog 1 (DLG

     1) in mammalian cells or tissues in vivo or in vitro.

18. A method of treating a disease associated with at least one Discs large homolog 1 (DLG 1) polynucleotide and/or at least one encoded product thereof, comprising:

administering to a patient a therapeutically effective dose of at least one antisense oligonucleotide of 12 to 30 nucleotides in length that binds and specifically hybridizes to a natural antisense sequence of said at least one Discs large homolog 1 (DLG

     1) polynucleotide and upregulates expression of said at least one Discs large homolog 1 (DLG1) polynucleotide;

thereby treating the disease associated with the at least one Discs large homolog 1 (DLG

     1) polynucleotide and/or at least one encoded product thereof.

19. The method of claim 18, wherein the disease associated with the at least one Discs large homolog 1(DLG 1) polynucleotide is selected from:

a disease or disorder associated with abnormal function and/or expression of DLG 1, cancer, a disease or condition associated with or characterized by cellular proliferation, a disease or disorder caused by tumorigenic human virus, a disease or disorder associated with cell fate specification, a disease or disorder associated with impaired cell suppression, ocular carcinogenesis, ductal carcinoma, cervical neoplasia, malignant fibrous histiocytoma, Charcot-Marie-Tooth disease.

20. The method according to claim wherein the natural antisense transcript has SEQ ID NO:

2.

21. A method of treating a disease or condition associated with cell distribution integrity or polarity comprising the administration of an oligonucleotide of about 12 to 30 nucleotides in length wherein said oligonucleotide targets and specifically hybridizes to and is at least eighty percent (80%) complementary to a natural antisense sequence of a Discs large homolog 1 (DLG 1) gene or isoform thereof to a patient in need of treatment thereof.

22. The method according to claim 21 wherein the natural antisense sequence is selected from SEQ ID NO. 2.

23. The method according to claim 21 wherein the disease or condition is selected from a disease, condition or wound associated with a tissue.